Therapeutic composition including saw palmetto for cancer treatment

ABSTRACT

A therapeutic herbal composition comprising Saw Palmetto, Bromelain, Willow Herb, Grape Seed Complex, Wild Rosella, Liquorice, Passionfruit Seed and Selenium Yeast is claimed. The composition may be used as a general health promotant but is also useful as a treatment of cancer and inflammation. Prostrate cancer and hyperplasia may be particularly beneficially addressed with the composition. The invention extends to a method of treatment using a composition as described.

FIELD OF THE INVENTION

[0001] This invention relates to a method for using natural plantproducts in the promotion of health and treatment or prevention ofdisease. The invention particularly relates to the use of a herbalformula of naturally occurring products, derivatives thereof orsynthetic equivalents for enhancing bodily, including prostatic health,and for the prevention or treatment of cancer, including prostaticcancer, and/or benign prostatic hyperplasia inflammation.

BACKGROUND OF THE INVENTION

[0002] The prostate is a secretory gland located in the malegenito-urinary tract. It primarily serves the purpose of providingsecretory fluids for supporting sperm viability. The prostate howeverhas a high incidence of age related pathology.

[0003] Prostatic cancer is a disease of global significance and is oneof the major causes of death in adult males. Even when non-fatal, thedisease can be intrusive, painful and disruptive to the lifestyle of asufferer. Benign prostatic hyperplasia is also a common occurrence withsignificant side effects.

[0004] Traditional methods of treatment of prostatic neoplasia and evenhypertrophy have involved the use of therapeutic agents such asanti-androgens including estrogens which have undesirable side effects.Additionally or alternatively, more radical means of treatment haveinvolved the application of surgery, radiation therapy and chemotherapy.While providing good results in some patients, these therapies have arelatively high rate of unpleasant side effects and an unacceptably highrate of recurrence of the targeted disease and associated clinicalsigns.

[0005] Some attempts have been made to use natural remedies for thetreatment of prostatic carcinoma. U.S. Pat. No. 5,666,5393 is directedto a composition comprising material from the herbs: Panaxpseudo-ginseng, Wall; Isatis indigotica, For; Ganoderma lucidum, Karst;Dendranthema morifolium, Tzvel; Glycyrrhzia glabra; Scutellariabaicalensis, Georgi; Rabdosia rubescens and Serenoa serrulata. Thisproduct is preferably provided in powdered concentrates from ethanolicextracts of dried plant material. The preparation has strong in vitrosupporting evidence for the blocking of cell replication in the prostatecell line PC-3 (Darzynkiewicz et al. 2000). Studies were conducted inwhich PSA levels were significantly reduced using the formula disclosedand in some cases PSA levels dropped to below the limit of detection.However, significant side effects were reported in several of thestudies (Small et al. 2000 and Oh et al. 2000). The side effectsincluded loss of libido (100%) and potency (100%),gynecomastia/gynecodemia (100%), diarrhoea (33%) and leg cramps (64%),while some 6% of subjects experienced deep vein thrombosis(Darzynkiewicz et al. 2000). These side effects are, at least in part,due to the estrogenic activity which has long been a form of therapy fortreatment of prostate pathologies. Serenoa serrulata has been welldocumented to contain phytoestrogens that lower endogenous estrogens inhost animals as well as inhibit proliferation of benign and cancerousprostatic cells (Carilla et al., 1984; Ichic et al., 1995; Delos et al.,1995).

[0006] It would be advantageous to produce a herbal tonic and remedywhich provides an alternative to known treatments, particularly forprostatic cancer and benign prostate hyperplasia, and which minimises orhas a reduced level of side effects during therapy.

[0007] Reference to any prior art in this specification is not andshould not be taken as an acknowledgment or any form of suggestion thatthis prior art forms part of the common general knowledge in anycountry.

SUMMARY OF THE INVENTION

[0008] Throughout this specification, unless the context requiresotherwise, the word “comprise”, or variations such as “comprises” or“comprising”, will be understood to imply the inclusion of a statedelement or integer or group of elements or integers but not theexclusion of any other element or integer or group of elements orintegers.

[0009] The bibliographic details of the publications referred to byauthor in this specification are collected at the end of thedescription.

[0010] The present invention is predicated in part on the identificationand combination of a number of plants or parts thereof which are usefulin the treatment or prophylaxis of cancer and/or inflammation, and inparticular, prostatic cancer, and in the promotion of well being andhealth in a subject.

[0011] Accordingly, a first aspect of the present invention contemplatesa herbal composition for the treatment or prophylaxis of a conditionassociated with neoplasia or a tendency to neoplasia, or for thepromotion of health and well being, the composition comprising Sawpalmetto, Bromelain, Willow herb, Grape seed complex, Wild rosella,Liquorice, Passionfruit seed and Selenium yeast.

[0012] In this context, Saw palmetto is Serenoa serrulata, Bromelain isor is derived from Ananas comosus. Willow herb includes Epilobiumparviflorum. Grape seed complex includes Vitis vinifera. Wild rosellaincludes Hibiscus sabdariffa. Liquorice includes Glycyrrihza glabra L.Passionfruit seed includes or is derived from Passiflora edulis andpreferably Passiflora edulis f. edulis. Selenium yeast is inactivatedwhole cell yeast product containing elevated levels of the essentialtrace element selenium in its natural food form.

[0013] In a preferred embodiment, the composition comprises ingredientsin the following ranges: Saw Palmetto   5-90%; Bromelain  1.5-21.5%;Willow Herb  0.5-14.5%; Grape Seed Complex 0.75-18.5%; Wild Rosella 0.5-20%; Liquorice  0.5-50%; Passionfruit Seed  0.5-20%; and SeleniumYeast  0.5-10%.

[0014] Reference in this specification to a plant either by way of itscommon name or species encompasses genetically modified plants.Genetically modified plants include transgenic plants or plants in whicha trait has been removed or wherein an endogenous gene sequence has beendown regulated, mutated or otherwise altered including the alteration orintroduction of genetic material which exhibits a regulatory effect on aparticular gene. Consequently, a plant which exhibits a characteristicnot naturally present in a species referred to is neverthelessencompassed by the present invention and is included within the scope ofthe above mentioned term.

[0015] Extracts of the plants may be used to produce the composition. Anextract may comprise sap or liquid or semi-liquid material exuded fromor present in leaves, stems, flowers, seeds, roots, bark or between thebark and the stems. An extract may therefore comprise liquid orsemi-liquid material located in fractions extracted from sap, leaves,stems, flowers, roots, bark or other plant material for example. Plantmaterial may even be subject to physical manipulation to disrupt plantfibres and extra cellular matrix material and inter and intra tissuesubject to extraction into a solvent including an aqueous environment.Alternatively, components of a plant may be mixed to form thecomposition of the invention. Those components may be dried and groundprior to mixture.

[0016] Reference herein to a plant includes various varieties, strains,lines, hybrids sub-species and variants or derivatives of the plant aswell as its botanical or horticultural relatives. Furthermore, thepresent invention may be practised using a whole plant or parts thereof,using sap or seeds or any other reproductive material which may be used.

[0017] In a further aspect, the invention resides in a method oftreating a subject, said method comprising the step of administering atherapeutically effective amount of a composition comprising Sawpalmetto, Bromelain, Willow herb, Grape seed complex, Wild rosella,Liquorice, Passionfruit seed and Selenium yeast. Preferably the methodis directed towards treatment of a mammal and most preferably a human.The method may be directed towards promoting well being or prostatehealth and/or preventing and/or treating prostatic cancer. The methodmay be directed to prevention or treatment of inflammation. The methodmay include the step of administering tablets/capsules to a subject. Atreating physician may select any regime that provides a beneficialeffect. However, a suitable regime may involve tablets/capsulesadministered once, twice or three times daily. A preferred regime is twoto three tablets/capsules twice daily or 2000 mg to 4000 mg daily ofcombined product.

[0018] By “effective amount”, in the context of modulating aninflammatory response or treating or preventing a disease or condition,is meant the administration of that amount of composition to anindividual in need thereof, either in a single dose or as part of aseries, that is effective for that modulation, treatment or prevention.The effective amount will vary depending upon the health and physicalcondition of the individual to be treated, the taxonomic group ofindividual to be treated, the formulation of the composition, theassessment of the medical situation, and other relevant factors. It isexpected that the amount will fall in a relatively broad range that canbe determined through routine trials.

[0019] In still a further aspect, the present invention resides in aherbal composition when used in the manufacture of a medicament for thetreatment or prevention of cancer or inflammation or otherwise promotinghealth the herbal composition comprising Saw palmetto, Bromelain, Willowherb, Grape seed complex, Wild rosella, Liquorice, Passionfruit seed andSelenium yeast.

[0020] In yet a further aspect, the invention contemplates use of acomposition as described above when administered in a symptomameliorating amount for a pathological condition and/or prophylaxis ofsaid pathological condition.

BRIEF DESCRIPTION OF DRAWINGS

[0021] 15 FIG. 1 is a schematic representation of a cell divisionalcycle.

[0022]FIG. 2 is a graphical representation of a control PC3 cell cycleassay.

[0023]FIG. 3 is a graphical representation of a PC3 cell cycle assay inthe presence of paclitaxol.

[0024]FIG. 4 is a graphical representation of a PC3 cell cycle assay inthe presence of a first level of a composition of the present invention.

[0025]FIG. 5 is a graphical representation of a PC3 cell cycle assay inthe presence of a second higher level of the composition of FIG. 4 ofthe present invention.

[0026]FIG. 6 is a graphical representation of an LNCap cell cycle assaywith a negative control.

[0027]FIG. 7 is a graphical representation of an LNCap cell cycle assaywith a positive control.

[0028]FIG. 8 is a graphical representation of an LNCap cell cycle assaywith a composition of the present invention.

[0029]FIG. 9 is a graphical representation of a HL60 cell cycle assaywith a negative control.

[0030]FIG. 10 is a graphical representation of a HL60 cell cycle assaywith a positive control.

[0031]FIG. 11 is a graphical representation of a HL60 cell cycle assaywith a composition of the present invention.

[0032]FIG. 12 is a graphical representation of the cytotoxic effect of acomposition of the present invention at different concentrations.

[0033]FIG. 13 is a graphical representation of percentage of cell growthinhibition plotted against the concentration of a composition of thepresent invention.

[0034]FIG. 14 is a graphical representation of the percentage of cellgrowth inhibition plotted against the concentration of the compositionof FIG. 13.

[0035]FIG. 15 shows the results of competitive binding of estradiol.

[0036]FIG. 16 is a chemical profile of a composition of the presentinvention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0037] The present invention is predicated in part on the identificationof biologically useful properties of a herbal formula comprisingnaturally occurring herbs and plants or their botanical or horticulturalrelatives. The biologically useful properties include their use in theprophylaxis and/or treatment of cancer or neoplasia, extend toanti-inflammatory activity and also to general promotion of health,particularly prostate health.

[0038] The term “treatment” is used in its broadest sense and includesthe prevention of neoplasia or cancerous cell growth, the inhibition ofgrowth or reduction in the size of a cancerous lesion as well asfacilitating the amelioration of the effects of cancer and/orinflammation and promotion of good health in a subject.

[0039] The term “prophylaxis” is also used herein in its broadest senseto encompass a reduction in the risk of development of cancer, aninflammatory condition or other condition. The prophylacticadministration of the composition may result in the composition becominginvolved in the treatment of the pathological condition. Use of theterms “treatment” or “prophylaxis” is not to be taken as limiting theintended result which is to reduce the incidence of cancer and inparticular prostatic cancer as well as or alternatively preventing orreducing inflammation or generally promoting health and well beingbenign prostatic hyperplasia.

[0040] Reference herein to a “subject” includes a human, primate,livestock animal (e.g. sheep, cow, horse, pig, goat, donkey), laboratorytest animal (e.g. mouse, rat, guinea pig, hamster), companion animal(e.g. dog, cat) or avian species such as poultry birds (e.g. chicken,ducks, turkeys, geese) or game birds (e.g. ducks, pheasants).

[0041] The preferred subject is a human or primate or laboratory testanimal.

[0042] The present composition is a balanced combination of six herbs,Selenium yeast and the natural enzyme Bromelain from pineapple. In thepreferred embodiment, the composition provides a formulation thatcombines safety, efficacy and ease of use for a subject undergoingtreatment with the invention.

[0043] Depending on the specific conditions being treated, thecomposition may be formulated and administered systemically or locally.Techniques for formulation and administration may be found in“Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa.,latest edition. Suitable routes may, for example, include oral, rectal,transmucosal, or intestinal administration; parenteral delivery,including intramuscular, subcutaneous, intramedullary injections, aswell as intrathecal, direct intraventricular, intravenous,intraperitoneal, intranasal, or intraocular injections. For injection,the composition of the invention may be formulated in aqueous solutions,preferably in physiologically compatible buffers such as Hanks'solution, Ringer's solution, or physiological saline buffer. Fortransmucosal administration, penetrants appropriate to the barrier to bepermeated are used in the formulation. Such penetrants are generallyknown in the art. Intra-muscular and subcutaneous injection may beappropriate.

[0044] The composition may be formulated readily using pharmaceuticallyacceptable carriers well known in the art into dosages suitable for oraladministration. Such carriers enable the compounds of the invention tobe formulated in dosage forms such as tablets, pills, capsules, liquids,gels, syrups, slurries, suspensions and the like, for oral ingestion bya patient to be treated. These carriers may be selected from sugars,starches, cellulose and its derivatives, malt, gelatine, talc, calciumsulphate, vegetable oils, synthetic oils, polyols, alginic acid,phosphate buffered solutions, emulsifiers, isotonic saline, andpyrogen-free water.

[0045] Pharmaceutical compositions suitable for use in the presentinvention include compositions wherein the active ingredients arecontained in an effective amount to achieve their intended purpose. Thedose of agent administered to a patient should be sufficient to effect abeneficial response in the patient over time such as a reduction in thesymptoms associated with the presence of a neoplasm or inflammation orto promote general well being and good health. The quantity of theagent(s) to be administered may depend on the subject to be treatedinclusive of the age, sex, weight and general health condition thereof.In this regard, precise amounts of the composition for administrationwill depend on the judgement of the practitioner. In determining theeffective amount of the chemical agent to be administered in thetreatment or prophylaxis of a condition, the physician may evaluatetissue or fluid levels of the biological entity, and progression of thedisorder. In any event, those of skill in the art may readily determinesuitable dosages of the compositions of the invention.

[0046] Pharmaceutical formulations for parenteral administration includeaqueous solutions of the herbal mixture in water-soluble form.Additionally, suspensions of the herbal mixture may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

[0047] Pharmaceutical preparations for oral use can be obtained bycombining the composition or active compounds thereof with solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose, and/or polyvinyl-pyrrolidone (PVP). If desired,disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate. Such compositions may be prepared by any of the methods ofpharmacy but all methods include the step of bringing into associationone or more chemical agents as described above with the carrier whichconstitutes one or more necessary ingredients. In general, thepharmaceutical compositions of the present invention may be manufacturedin a manner that is itself known, e.g. by means of conventional mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping or lyophilising processes.

[0048] Dragee cores are provided with suitable coatings. For thispurpose, concentrated sugar solutions may be used, which may optionallycontain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel,polyethylene glycol, and/or titanium dioxide, lacquer solutions, andsuitable organic solvents or solvent mixtures. Dyestuffs or pigments maybe added to the tablets or dragee coatings for identification or tocharacterize different combinations of active compound doses.

[0049] Pharmaceutical compositions which can be used orally includepush-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added.

[0050] Dosage forms of the invention may also include injecting orimplanting controlled releasing devices designed specifically for thispurpose or other forms of implants modified to act additionally in thisfashion. Controlled release of an agent of the invention may be effectedby coating the same, for example, with hydrophobic polymers includingacrylic resins, waxes, higher aliphatic alcohols, polylactic andpolyglycolic acids and certain cellulose derivatives such ashydroxypropylmethyl cellulose. In addition, controlled release may beeffected by using other polymer matrices, liposomes and/or microspheres.

[0051] The herbal components of the invention may be provided as saltswith pharmaceutically compatible counterions. Pharmaceuticallycompatible salts may be formed with many acids, including but notlimited to hydrochloric, sulphuric, acetic, lactic, tartaric, malic,succinic, etc. Salts tend to be more soluble in aqueous or otherprotonic solvents that are the corresponding free base forms.

[0052] For any composition used in the method of the invention, thetherapeutically effective dose can be estimated initially from cellculture assays. For example, a dose can be formulated in animal modelsto achieve a circulating concentration range that includes the IC₅₀ asdetermined in cell culture. Such information can be used to moreaccurately determine useful doses in humans.

[0053] Toxicity and therapeutic efficacy of such chemical agents can bedetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, e.g. for determining the LD₅₀ (the dose lethal to50% of the population) and the ED₅₀ (the dose therapeutically effectivein 50% of the population). The dose ratio between toxic and therapeuticeffects is the therapeutic index and it can be expressed as the ratioLD₅₀/ED₅₀. Compositions that exhibit large therapeutic indices arepreferred. The data obtained from these cell culture assays and animalstudies can be used in formulating a range of dosages for use in humans.The dosage of such compounds lies preferably within a range ofcirculating concentrations that include the ED₅₀ with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized. The exactformulation, route of administration and dosage can be chosen by theindividual physician in view of the patient's condition (see for exampleFingl et al., 1975, in “The Pharmacological Basis of Therapeutics”, Ch.1 μl).

[0054] Dosage amount and interval may be adjusted individually toprovide plasma levels of the active agents which are sufficient tomaintain symptom-ameliorating effects. Usual patient dosages forsystemic administration range from 1-5000 mg/day, commonly from 1-3000mg/day. Stated in terms of patient body weight, usual dosages range from0.02-80 mg/kg/day, commonly from 0.02-50 mg/kg/day, typically from0.2-50 mg/kg/day. Stated in terms of patient body surface areas, usualdosages range from 0.5-2500 mg/m²/day, commonly from 0.5-1500 mg/m²/day,typically from 5-1500 mg/m²/day.

[0055] Alternatively, one may administer the compound in a local ratherthan systemic manner, for example, via injection of the compounddirectly into a tissue, often in a depot or sustained releaseformulation. Furthermore, one may administer the drug in a targeted drugdelivery system, for example, in a liposome coated with tissue-specificantibody. The liposomes will be targeted to and taken up selectively bythe tissue. In cases of local administration or selective uptake, theeffective local concentration of the agent may not be related to plasmaconcentration.

[0056] The herbal composition of the invention can also be deliveredtopically. For topical administration, a composition containing between0.001-5% or more composition is generally suitable. Regions for topicaladministration include the skin surface and also mucous membrane tissuesof the vagina, rectum, nose, mouth, and throat. Compositions for topicaladministration via the skin and mucous membranes should not give rise tosigns of irritation, such as swelling or redness.

[0057] The topical composition may include a pharmaceutically acceptablecarrier adapted for topical administration. Thus, the composition maytake the form of a suspension, solution, ointment, lotion, sexuallubricant, cream, foam, aerosol, spray, suppository, implant, inhalant,tablet, capsule, dry powder, syrup, balm or lozenge, for example.Methods for preparing such compositions are well known in thepharmaceutical industry.

[0058] In one embodiment, the composition may be administered topicallyto a subject, e.g. by the direct laying on or spreading of thecomposition on the epidermal or epithelial tissue of the subject, ortransdermally via a “patch”. Such compositions include, for example,lotions, creams, solutions, gels and solids. Suitable carriers fortopical administration preferably remain in place on the skin as acontinuous film, and resist being removed by perspiration or immersionin water. Generally, the carrier is organic in nature and capable ofhaving dispersed or dissolved therein a chemical agent of the invention.The carrier may include pharmaceutically-acceptable emolients,emulsifiers, thickening agents, solvents and the like.

[0059] A preferred formulation of the composition of the invention isprepared to provide a balanced combination of Saw palmetto (Serenoaserrulata), the natural enzyme bromelain from pineapple and six herbsincluding Willow herb (Epilobium parviflorum), Grape seed complex (Vitisvinifera), Wild rosella (Hibiscus sabdariffa), Liquorice (Glycyrrihzaglabra L.), Passionfruit seed (Passiflora edulis f. edulis) and Seleniumyeast. The components are preferably utilised in the dried powder form.

[0060] A composition of the herbal formula is preferably presented as atablet/capsule which may have approximately 500˜800 mg of activeingredients per tablet/capsule. Examples of typical formulation ofactive ingredients may be produced with percentage active ingredient ona weight basis as set out in Table 1: TABLE 1 Twenty-five formulas ofrepresentative compositions of the present invention Summaries (VALUESIN PERCENTAGES) SAW WILLOW GRAPE SEED PASSION SELENIUM PALMETTOBROMELAIN HERB COMPLEX WILD ROSELLA LIQUORICE FRUIT YEAST 1 90.00 4.502.25 .75 .50 .50 .50 1.00 2 85.00 1.50 4.50 4.50 .50 2.00 1.00 1.00 380.00 8.50 .50 1.50 2.50 2.50 .50 4.00 4 75.00 8.50 1.00 1.00 1.00 .503.00 10.00 5 28.20 10.60 2.80 2.80 1.40 49.30 1.40 3.50 6 38.80 10.602.80 2.80 2.20 37.20 1.60 4.00 7 48.50 10.60 3.80 2.80 2.10 25.00 2.205.00 8 20.20 12.60 5.80 13.80 12.80 23.80 4.50 6.50 9 15.50 15.00 8.8010.60 20.00 15.00 12.60 2.50 10 10.50 15.00 9.50 16.50 12.50 18.50 15.002.50 11 5.00 15.00 9.50 18.50 14.50 19.50 16.00 2.00 12 10.00 12.00 5.5016.50 15.50 15.50 20.00 5.00 13 55.00 12.50 6.50 5.20 7.80 5.20 5.002.80 14 41.00 13.00 7.50 4.50 12.50 9.50 6.50 5.50 15 35.50 18.50 13.504.00 12.00 8.50 5.50 2.50 16 30.50 20.50 10.00 5.50 5.50 12.50 10.505.00 17 25.00 21.50 14.50 5.50 6.50 11.50 11.50 4.00 18 65.00 10.00 5.005.00 5.00 3.00 4.00 3.00 19 61.50 5.50 6.50 10.50 3.20 4.50 3.50 4.80 2045.50 18.50 5.50 11.50 6.50 7.50 4.50 .50 21 32.00 13.50 9.50 16.5010.50 8.50 4.50 5.00 21 32.00 13.50 9.50 16.50 10.50 8.50 4.50 5.00 2270.00 5.00 5.00 5.00 5.00 3.50 3.00 3.50 22 70.00 5.00 5.00 5.00 5.003.50 3.00 3.50 23 50.00 8.00 2.50 2.50 5.50 25.50 2.50 3.50 23 50.008.00 2.50 2.50 5.50 25.50 2.50 3.50 24 42.50 10.50 4.50 4.50 2.50 30.503.00 2.00 24 42.50 10.50 4.50 4.50 2.50 30.50 3.00 2.00 25 72.00 9.503.00 4.00 3.50 5.00 1.50 1.50 25 72.00 9.50 3.00 4.00 3.50 5.00 1.501.50

[0061] It is important to note that the above ratios and percentages areapproximate only. The components of the herbal formula provided in Table1 are natural products.

[0062] As such they are prone to considerable variation in strength dueto a wide range of influences including climatic, geographical andgenetic conditions. The concentration of the ingredients in thetherapeutic composition of the invention may be beneficially adjusted toaccommodate variation in activity in particular ingredients.

[0063] The preferred range of components are therefore Saw Palmetto5-90%; Bromelain 1.5-21.5%, Willow Herb 0.5-14.5%, Grape Seed Complex0.75-18.5%; Wild Rosella 0.5-20%; Liquorice 0.5-50%; Passionfruit Seed0.5-20% and Selenium yeast 0.5-10% calculated on a weight basis.

[0064] A tablet may be formed with additional components which mayinclude excipients and binders as used in the pharmaceutical industry.

[0065] While the following discussion is directed to anti-cancertherapy, it should be understood this is not limiting. The compositionmay be used as a general tonic and promoter of good health, inparticular, prostate gland health. Further the application in cancerprophylaxis and treatment may extend beyond prostatic neoplasia andinclude a range of tumours including sarcomas and carcinomas of otherorgans and sites.

[0066] The present composition has distinct advantages over some priorart remedies and therapies both in terms of its efficacy and decreasedlevel of side effects. Referring to FIG. 1, there is seen a schematicrepresentation of a cell cycle 10 (adapted from Voet & Praft 1999). Theprocess by which cells divide and DNA is replicated is a complicatedprocess in eukaryotic cells and involves a number of different phases.Cancerous cells are dividing cells and so will exhibit a well definedcell cycle which is separated into several distinct phases. It starts atthe G₁ phase 11 or the first gap phase. If the cells are permanentlyarrested in G₁, as in non-dividing cells, it is then called G₀ 12. At G₁the cell contains two copies of each chromosome. As the cell cycleprogresses from G₁, it enters the synthesis or S phase 13 and duringthis phase DNA is replicated. When this replication is finished, thecell enters the G₂ phase or the second gap phase 14. At the end of theG₂ phase the cell is ready to enter the M phase or mitotic phase 15during which the cell divides. Using cell cycle analysis it is possibleto break down the events into the major phases such as G₀-G₁, G₂-M andS. Many drugs used in the fight against cancer affect the cell cycle atthese points.

EXAMPLE 1

[0067] Cell cycle analysis was carried out using tablets/capsules of thepresent invention. The formulation was tested as a single tablet/capsuleconcentration in triplicate and as a two capsule concentration also intriplicate. The effects on the cell cycles at various concentrationswere therefore comparable.

[0068] Tablets/capsules were weighed, crushed and extracted with 10 mlof methanol. The extracts were sonicated for 40 mins and then spun downat 4000 rpm. The supernatant was removed and 100 μL of this extract wasused in the cell cycle analysis. A further aliquot of this extract wassubjected to HPLC-MS analysis as described in Example 3 below. Cellswere split into 16×25 cm² flasks at a low concentration and allowed togrow to 65-70% confluence (at the time of addition the cells were in logphase). Flasks were incubated for 24 hours and control flasks of controlmedia (no additions), an ethanol (100 μL EtOH), a control methanol (100μL MeOH) and paclitaxol (10 μL of 0.01 mg/mL=10 ng/mL) were also run.

[0069] Flow cytometry was performed on the fixed cells after washing inPBS and stained using propidium iodide (2% PI in 0.1% Triton X-100containing 2 mg/mL Rnase A). A Becton Dickson FACSCaliber was used toassay the cells.

[0070] Cell cycle analysis using flow cytometry of propidium iodidestained cells showed the tablets affect the cell cycle of PC-3 celllines by arresting the cell cycle in G₂-M.

[0071] Even a low dose of the invention preferentially arrested thecells in G₂-M. While the cell line is a prostate cancer cell line it ispossible to apply the same therapy to other forms of cancer cell linesand therefore to other cancers.

[0072] The following table sets out the results of the test whereinformulations of the present invention are identified as Herb Formula:TABLE 2 Percentage Changes in G₀-G₁ and G₂-M G₀-G₁ G₂-M Control 43.7425.43 Herb Formula-1-1 14.08 75.35 Herb Formula-1-2  7.88 74.92 HerbFormula-1-3 18.08 55.39 Mean ± SD 13.35 ± 5.14 68.55 ± 11.40 HerbFormula-2-1  4.45 82.12 Herb Formula-2-2 20.91 63.88 Herb Formula-2-3 8.76 74.53 Mean ± SD 11.37 ± 8.54 73.51 ± 9.16 

[0073] It is apparent in Table 2 that the result for the control showsthat only 44% of the cells are in the G₀-G₁ phase while 25% are in theG₂-M phase. With increasing dosage of the tablet/capsule of theinvention, the mean percentage of cells in the G₀-G₁ was in the range of13.35 to 11.37%. Meanwhile the percentage of cells in the G₂-M phase hadrisen to be between on average 68.55-73.51%. This indicates a locking orarresting of cells in the G₂-M stage and subsequent prevention ofmitotic and cellular growth. FIG. 2 is a histogram of the results offlow cytometry performed on the control. In this result 43.74% of thecells were in the G₀-G₁ phase at 50.80, 25.43% were in the G₂-M phase at99.88. 30.83% of the cells were in the S phase.

[0074]FIG. 3 shows the results when using Paclitaxol, a well knownchemotherapeutic agent for use against cancer. In this result, 1.16% ofthe cells are in the G₀-G₁ phase at 25.06, 71.89% of the cells in theG₂-M phase at 98.40. This highlights the fact that the cells arearrested at the G₂-M phase with this drug. 26.95% were in the S phase.

[0075]FIG. 4 shows the results of using one tablet/capsule of thepresent invention. 18.08% of the cells were in the G₀-G₁ phase at 48.12.55.39% of the cells were in the G₂-M phase at 95.52 while 26.53% were inthe S phase.

[0076]FIG. 5 shows the results of using two tablets/capsules of thepresent invention. The result here is even more marked in that only4.45% of the cells were in the G₀-G₁ phase at 53.86. Meanwhile 82.12% ofthe cells were in the G₂-M phase at 105.22 and 13.43% of the cells werein the S phase.

[0077] These results highlight the therapeutic effectiveness of thepresent invention against replication of prostatic carcinoma cellslines. Tests have also shown that in clinical use, this product mayproduce lowering of prostatic specific antigen (PSA) which is a wellknown indicator of prostatic hyperplasia and prostatic carcinoma.

[0078] The present invention has also been shown to have ananti-inflammatory effect and may be of use in inflammatory conditionssuch as arthritis and autoimmune diseases. The invention may also beuseful against secondary inflammation produced as a result of primaryinfectious or traumatic processes.

EXAMPLE 2

[0079] A anti-cancer activity assay was conducted with the resultspresented in Table 3 wherein the herb formula represents a combinationaccording to the present invention. LNCap is an androgen-dependentprostate cancer cell line.

[0080] From cell cycle analysis using flow cytometry of promidium iodidestained cells 3.2 and 1.6 mg/mL concentrations of the herb formulaaffect the cell cycle of LNCap cell lines by arresting the cell cycle inG₂-M. The herb formula showed dose-dependent effects on LNCap cell linesassay. TABLE 3 Herb Formula bioactivity using LNCap cell cycle assayConcentration Treatment (mg/10 mL) G₀-G₁ G₂-M Negative control 80.795.31 Herb Formula 0.4 62.72 10.83 0.8 72.23 9.91 1.6 10.88 79.69 3.210.73 77.23

[0081]FIG. 6 is a graphical representation of the DNA histogram of aLNCap cell line with a negative control.

[0082]FIG. 7 is a graphical representation of an LNCap cell cycle assaywith a positive control in the form of Paclitaxol showing arresting ofcells in G₂-M.

[0083]FIG. 8 shows the results of a combination of the presentinvention, the herb formula composition in contact with an LNCap cellcyle assay. As noted, the herb formula shows a dose dependent effect onLNCap cell lines and arrests the cells in the G₂-M stage.

EXAMPLE 3

[0084] The bioactivity of the herb formula was assessed using HL60 celllines. HL60 is a human promyelocytic leukemia cell line.

[0085] From cell cycle analysis using flow cytometry of promidium iodidestained cells 3.2 and 1.6 mg/mL concentrations of the herb formulaaffect the cell cycle of HL60 cell lines by arresting the cell cycle inG₂-M. Herb Formula showed dose-dependent effects on HL60 cell linesassay as shown in Table 4. TABLE 4 The herb formula bioactivity usingHL60 cell cycle assay Concentration Treatment (mg/10 mL) G₀-G₁ G₂-MNegative control 34.79 12.77 Herb Formula 0.4 31.65 13.74 0.8 32.4412.07 1.6 28.65 21.89 3.2 12.34 52.68

[0086]FIG. 9 shows a histogram of cell cycle analysis for an HL60 cellline with a negative control.

[0087]FIG. 10 shows a graphical representation of the DNA histogram foran HL60 cell line with a positive control in the form of Paclitaxol. Theeffect is to arrest the cells in the G₂-M stage.

[0088]FIG. 11 shows a graphical representation of the DNA histogram foran HL60 cell line with the herb formula. The mixture displayschemotherapeutic benefit by arresting the cells in the G₂-M stage.

EXAMPLE 4

[0089] As noted, the herbal formula may be used as an anti-inflammatoryagent. This capacity was investigated with a bioactivity assay

[0090] This assay was run to determine the effectiveness of the herbalformula to inhibit Prostaglandin E₂ (PGE₂) production in a mouse 3T3fibroblast cell line.

[0091] The anti-inflammatory assay measures the level of PGE₂ instimulated mouse fibroblast 3T3 cell line after treatment with theherbal formula. The use of a stimulated cell line rather than usingisolated enzyme systems gives more useful data.

[0092] PGE₂ is formed in a variety of cells from PGH₂, which itself issynthesized from arachiodonic acid by the enzyme prostaglandinsynthetase. The assay measures secreted levels of PGE₂ from thesupernatant of the fibroblast cells.

[0093] The herbal formula had significant dose dependent inhibition ofsecreted PGE₂ from the mouse fibroblast cell line equivalent to orbetter than aspirin under the test conditions. TABLE 5 Percentinhibition of secreted PGE₂ from 3T3 cells exposed to the herb formulaConcentration Sample Dilution (μg/mL) % Control Aspirin 500 500 μM 14.2Herb Formula 500 1000 15.8 Herb Formula 500  100 30.0

EXAMPLE 5

[0094] Safety and toxicity of the herbal formula were investigated usinga cytotoxicity assay.

[0095] Cells were split into 10 cm² tissue culture tubes at a lowconcentration and allowed to grow to confluent. Media was DMEM with 10%horse sera (GIBCO) with glutamine and pen/strep added tubes placed in a10% CO₂ incubator. Cells were then transferred to tissue culture platesin 99 μL media. Samples for testing were then added, 1 μL into each wellranging from 1 mg/mL to 0.001 mg/mL. The assay is based on the presenceof ATP in living cells and is measured using luminescence readings onthe Wallac Microbeta.

[0096] The herb formula only had slight cytotoxic affect at the 0.1mg/mL doses and almost no affect at the 0.01 and 0.001 mg/mL dose asshown in FIG. 12.

EXAMPLE 6

[0097] The toxicity of the herbal formula was also investigated usingHL60 cell lines. The herbal formula had cytotoxic affect at the 100μg/mL doses, had a slight cytotoxic effect at the 33 μg/mL dose and hadno affect at the 3.7 and 1.12 μg/mL dose. The results are shown in FIG.13 which is a chart of percentage cell growth inhibition againstconcentration of Herbal Formula.

EXAMPLE 7

[0098] The toxicity of the herbal formula was also investigated usingHepG2 cell lines. Unlike the other cell lines, the herbal formula hadslight cytotoxic affect at the 300 μg/mL doses and had no affect at the100, 33, 11.1, 3.7 and 1.12 μg/mL dose.

[0099] The results are shown in FIG. 14 which is a chart of percentagecell growth inhibition against concentration of the herb formula.

EXAMPLE 8

[0100] The estrogen binding capacity of the herbal formula wasinvestigated by assay. This assay was developed and run to determine ifthe herbal extracts that constitute the herbal formula have any bindingactivity towards the estrogen (E₂) binding site on MCF-7 cell lines. TheMCF-7 is a breast cancer cell line and was chosen because of the abilityto express E₂ binding sites.

[0101]³H 17-β-estradiol (E₂) was used at a final concentration in thewells of 1 nM in media. Samples were incubated for 3 hours after whichthe wells were aspirated and washed 3 times with warm PBS. Cell boundradioactivity was extracted with 300 μL EtOH which was added to eachwell and left to lyse for 20 min at room temperature. An aliquot wasremoved for counting, transferred to a counting plate and scintillantadded and the plate counted in a Perkin Elmer MicroBeta.Diethylstilbestrol (DES) (100 μM) and diadzein (197 μM) were used aspositive binding controls. Cold 17-β-estradiol (non-radioactive)(0.051-12.5 nM) was used as a competitor for the E₂ binding sites. Acompetitive binding of ³H 17-β-estradiol against cold 17-β-estradiol wasdemonstrated by this assay, the results of which are shown in FIG. 15.As expected both DES (100 μM) and diadzein (197 μM) bind to the E₂receptor displacing over 85% and 60% of the ³H E₂, respectively.

[0102] Herb Formula however has less than 30% inhibition of ³H17-β-estradiol binding to E₂ receptors at the highest concentrationtested (100 μg/mL). The other concentrations of the herb formula hadlittle or no effect on the inhibition of ³H 17-β-estradiol binding. Fromthese results it can not be predicted with certainty whether or not thecomposition will have estrogenic activity in whole animals, it canhowever be inferred that since there is little or no binding to the E₂receptors (at the concentrations tested) it would seem unlikely thatestrogenic activity could occur. This would also suggest that the herbformula may well be free of associated disturbing side-affects (breasttenderness, loss of libido etc).

EXAMPLE 9

[0103] Toxicity of the present invention was assessed using P₃₈₈ cellline toxicity assay. Cells were split into 10 cm² tissue culture tubesat a low concentration and allowed to grow until confluent. The mediaused was DMEM with 10% horse sera (GIBCO) with glutamine and PEN-STREPadded tubes placed in a 10% CO₂ incubator. Cells were then transferredto tissue culture plates in 99 μL media. Samples for testing were thenadded (1 μL into each well ranging from 1 mg/ml to 0.001 mg/ml). Theassay was based on the presence of ATP in living cells and was measuredusing luminescence readings on the Wallac Microbeta. Tablets of thepresent invention showed a level of cytotoxity at the higher dose of 1mg/ml. However, it had only slight cytotoxic effect at the 0.1 mg/mldose and almost no effect at 0.01 and 0.001 mg/ml doses. This lack ofcytotoxicity shows a particular advantage for the present invention asin minimises risk to healthy tissues.

[0104] Although any suitable route of administration may be employed forproviding the patient with an effective dosage of the compositionaccording to the present invention, oral administration is preferred.Suitable alternative routes include, for example, oral, rectal,parenteral, intravenous, topical, transdermal, subcutaneous, intramuscular and similar forms of administration. Suitable dosage formsinclude tablets, troches, dispersions, suspensions, solutions, capsules,patches, suppositories and the like although oral dosage forms arepreferred.

EXAMPLE 10

[0105] The chemical profile of the extract prepared as outlined inExample 1 was obtained using an Agilent 1100 HPLC coupled to a diodearray detector and mass spectrometer. Separation was achieved on a 5 μmC₁₈ column (15 cm×4.6 cm; Phenomonex) using a linear gradient from water(95%) to acetonitrile (95%) over forty minutes at a flow rate of 1mL/min. A typical fingerprint of this invention should display thecharacteristic peak profile displayed in FIG. 16. The chromatograms inthis example were generated at 330 nm and 210 nm from the diode arraydata.

EXAMPLE 11

[0106] A white male, age 74, presented for a biopsy and bone scan whichrevealed prostate cancer without bony metastases in 1998. Serum PSA wasmeasured to be 300. This patient then started hormone therapy includingan oral administration of 100 mg of Androcur (Cyproterone Acetate) 3times per day, and a subcutaneous injection of 3.6 mg of Zolodex(Goserelin Acetate Implant) once per three months. The prostate has beenmonitored since with serum PSA levels, varying between “28-62”.

[0107] In May 2001, the PSA level was measured to be 41. At that time,the patient stopped taking Androcur and Zolodex, and started to takeCasodex 50 mg once daily. The treatment with Casodex continued for threemonths and follow-up blood tests indicated a serum PSA of 5.7.

[0108] From August 2001, this patient started to combine Casodex with anoral administration of the herbal formula of the present invention. Theintake dosage of the herbal formula was 2070 mg per day. Thiscombination treatment continued for two months. During the two months,the patient's energy and well-being were enhanced, and no adverseeffects were reported.

[0109] From October 2001, the dosage of this herb formula was increasedto 2760 mg per day. Following PSA level measurement was 2.3 in November2001. One month later (December 2001) blood tests showed a constant PSAreading of 2.3.

EXAMPLE 12

[0110] The patient in this case study was a 63-year-old white male. Abiopsy was performed in 1996, and the pathology report had indicatedBenign Prostatic Hyperplasia. The PSA was measured to be 21.

[0111] The prostate has been monitored approximately every 6 months. Asthe PSA reading was between “13-17” on occasions, when readings werehigh, antibiotics were prescribed for prostatitis. The PSA was measuredto be 18.7 at August of 2001.

[0112] Biopsy was performed again in November of 2001 and nine sampleswere taken in the biopsy and cancer was detected in one sample only. Theurologist suggested a “watchful waiting” position should be adopted andthen recommended a discussion with an Oncologist. The patient didcontact an Oncologist, and radiation therapy was suggested.

[0113] The patient did not take radiotherapy, and started a therapy byoral administration of the herbal formula of the present invention. Theintake dosage of herbal formula was 2760 mg per day. This combinationtreatment continued for one and a half months and a follow-up serum PSAof 18.1 was reported. During this period, the patient's energy andwell-being were enhanced. No adverse effects were reported.

EXAMPLE 13

[0114] A white male, age 58, presented a PSA level of 8.6. One weeklater, this patient started to take therapy by oral administration ofthe herbal formula of the present invention. The intake dosage of theherbal formula was 1380 mg per day. This patient did not take anyhormone therapy.

[0115] This herbal formula continued for one month, and a follow-upserum PSA of 3.7 was reported. During this period, the patient's energyand well-being were enhanced and no adverse effects were reported. Thispatient increased the dosage to 2760 mg per day. Four months later, thefollow-up serum PSA level was reported to be 2.1.

EXAMPLE 14

[0116] A white male, age 69, presented for a PSA level of 4.4 in July1994. This patient was put in a “watchful position”, and took two herbmixtures “Detoxitea” and “Essiac”. Six years later (July, 2000), theserum PSA level was reported to be 6.4. Then this patient added somenatural supplements including Epilobium and liquid selenium. The serumPSA level has been monitored regularly since that time as displayed infollowing table. TABLE 6 Serum PSA level Time Serum PSA (μg/ L) Ref.Range (μg/L)  5/7/94 4.4 0.0-4.0 17/7/00 6.4 * 15/11/00 7.7 * 17/01/017.6 * 12/06/01 8.8 * 17/09/01 10.9 * 17/10/01 11.2 * 17/11/01 11.5 *30/11/01 Start Herb Formula 15/01/02 10.6 *

[0117] From the end of November 2001, the patient started therapy byoral administration of the herbal formula of the present invention. Theintake dosage of the herbal formula was 2760 mg per day. This herbaltreatment continued for one and half months, a follow-up serum PSA levelwas 10.6. During the two months, the patient's energy and well-beingwere enhanced, and no adverse effects were reported.

[0118] Throughout the specification the aim has been to describe thepreferred embodiments of the invention without limiting the invention toany one embodiment or specific collection of features. Those of skill inthe art will therefore appreciate that, in light of the instantdisclosure, various modifications and changes can be made in theparticular embodiments exemplified without departing from the scope ofthe present invention. All such modifications and changes are intendedto be included within the scope of the disclosure.

BIBLIOGRAPHY

[0119] Carilla, E., Briley, M. Fauran, Fig. Sultan, C. and Duvilliers,C. (1984). Serenoa repens (Permixon), a new treatment for prostaticbenign hyperplasia, to the cytosolic andrgoen receptor in the ratprostate. J Steroid Biochem., 20, 521-581.

[0120] Darzynkiewicz, Z., Tragnaos, F, Wu, J. M. and Chen, S. (2000).Chinese herbal mixture PC SPES in treatment of prostate cancer, Int. J.Oncol., 17, 729-736.

[0121] Delos, S., Cargol, J. L., Ghazarossian, E. Raynaud J. and martin,P. M. (1995). Testosterone metabolism in primary cultures of humanprostate epithelial cells and fibroblasts. J Steroid Biochem. Mol.Biol., 55, 375-383.

[0122] Ichic, C., Delos, S., Guiro, O. Tate, R. Raynaud, J and Martin,P. M. (1995). Human prostatic steroid 5a-rediuctase isoforms—acomparative study of selective inhibitors. J. Steroid Biochem. Mol.Biol., 54, 237-79.

[0123] Kameda et. al. (1999). Annual meeting of the America Society ofClinical Oncologists, Abstract 1230.

[0124] Oh et al. (2000). Annual meeting of the America Society ofClinical Oncologists, Abstract 1334.

[0125] Pfeiffer, B. L., Pirani, J. F., Hamann, S. R. and Klippel, K.Fig. (2000). PC—SPES. A dietary supplement in the treatment ofhormone-refractory prostate cancer, BJU Int, 85, 481-485.

[0126] Small et. al, (2000). Annual meeting of the America Society ofClinical Oncologists, Abstract 1316.

[0127] Voet D, Voet J. G. & Pratt, C. W. (1999). “Fundamentals ofBiochemistry”, Eds Wiley & Sons, New York, p915.

What is claimed is:
 1. A therapeutic herbal composition comprising SawPalmetto, Bromelain, Willow Herb, Grape Seed Complex, Wild Rosella,Liquorice, Passionfruit Seed and Selenium yeast.
 2. The therapeuticherbal composition of claim 1 wherein Saw Palmetto is present in aconcentration in a range of 5-90% calculated by weight
 3. Thetherapeutic composition of claim 1 wherein the Bromelain is present in arange of between 1.5-21.5% calculated by weight.
 4. The therapeuticherbal composition of claim 1 wherein the Willow Herb is present in arange of between 0.5-14.5% calculated by weight.
 5. The therapeuticherbal composition of claim 1 wherein the grape seed complex is presentin a range of 0.75-18.5% calculated by weight.
 6. The therapeutic herbalcomposition of claim 1 wherein the wild rosella is present in a range of0.5-20% calculated by weight.
 7. The therapeutic herbal composition ofclaim 1 wherein the liquorice is present in a range of 0.5-50%calculated by weight.
 8. The therapeutic herbal composition of claim 1wherein the passionfruit seed is present in a range of 0.5-20%calculated by weight.
 9. The therapeutic herbal composition of claim 1wherein the selenium yeast is present in a range of 0.5-10% calculatedby weight.
 10. The therapeutic herbal composition of claim 1 when usedfor the treatment or prophylaxis of cancer in a subject.
 11. Thetherapeutic herbal composition of claim 10 wherein the cancer isprostate cancer.
 12. The therapeutic herbal composition of claim 1 whenused to prevent or limit inflammation in a subject.
 13. The therapeuticherbal composition of claim 1 when used to prevent or treat prostatichyperplasia in a subject.
 14. The therapeutic herbal composition ofclaim 1 when used as a health promotant in a subject.
 15. Thetherapeutic herbal composition of claim 1 when used to promote prostatehealth in a subject.
 16. The therapeutic herbal composition of claim 10wherein the subject is a human.
 17. A combination of Saw Palmetto,Bromelain, Willow Herb, grape seed complex, wild rosella, liquorice,passionfruit seed and selenium yeast when used for the production of amedicament for the treatment or prevention of a disease or for thepromotion of health in a subject.
 18. The combination of claim 17wherein the disease is associated with the prostate.
 19. The combinationof claim 17 wherein the subject is a human.
 20. A method of medicating asubject to prevent or treat disease and/or promote health, said methodcomprising the step of administering a therapeutically effectivecombination of Saw palmetto, Bromelain, Willow herb, Grape seed complex,Wild rosella, Liquorice, Passionfruit seed and Selenium yeast.
 21. Themethod of claim 21 wherein the combination comprises Saw Palmetto 5-90%;Bromelain 1.5-21.5%; Willow Herb 0.5-14.5%; Grape Seed Complex0.7518.5%; Wild Rosella 0.5-20%; Liquorice 0.5-50%; Passionfruit Seed0.5-20%; and Selenium Yeast 0.5-10% calculated by weight.